Storage and/or transportation of produce, flowers and plants

ABSTRACT

The disclosure relates to methods and compositions for improving storage and/or transportation of food, flowers, plants, or the like.

FIELD

The disclosure relates to methods and compositions for improving storageand/or transportation of food, flowers, plants, or the like.

BACKGROUND

Each year, millions of dollars of food, flowers, plants, or the like arelost to damage during transportation from harvest to consumers.Post-harvest preservation of foods and plants, particularly produce,flowers, and plants poses major technical challenges due to the fragilenature of the products and their susceptibility to changes intemperature, humidity, light, atmosphere, and other environmentalfactors. There is an unmet need for methods that allow for themaximization of the storage volume in warehouses, stores, and withintrucks and other systems that transport food, flowers, plants, or thelike from harvest to the point of sale and consumers in an economicallyfeasible and environmentally conscientious strategy.

SUMMARY

The presently described technology and its advantages will be betterunderstood by reference to the following embodiments, aspects, andexamples. These examples are provided to describe specific embodimentsof the present technology. Those with ordinary skill in the art willappreciate that various modifications and alternatives could bedeveloped in light of the overall teachings of the disclosure.Accordingly, the particular arrangements and examples disclosed aremeant to be illustrative only and not limiting as to the scope of theinvention, which is to be given the full breadth of the appended claims,and any and all equivalents thereof.

The disclosure provides methods and compositions that improve thepost-harvest preservation of food for human consumption (e.g., edibleprotein, processed food, meat, fish, shellfish, produce, vegetables,fruits, or the like), food for animal consumption (e.g., pet food,animal feed, or the like), flowers, plants, seeds, nuts, or the like(herein referred to as the “product”) and maximize the capacity oftransport systems that carry them from harvest to the point of sale andconsumers. The products may be coated by coatings that extend the shelflife, improve transport, protect product quality and freshness, and/orsafety. The coatings include edible and non-edible coatings.

In one embodiment, the disclosure provides a method for increasing thevolume and/or weight capacity of a system that stores and/or transportsproduct, wherein at least part of the surface of the product is coatedwith silk fibroin, chitin, mono- and di-acyl glycerides, shellac,starch, high fructose corn syrup, mayauba wax, maydelilla wax, beeswaxor other waxes, vegetable oil, paraffin oil, Ethylenediaminetetraaceticacid (“EDTA”), cellulose, pectin, alginate, chitosan, gum Arabic, soyprotein, zein, casein, and/or whey. In one embodiment, the system is atruck, a boat, an airplane, a transport and/or storage container, andthe like. In one embodiment, the system is refrigerated. In oneembodiment, the system is not refrigerated and carries the product atambient temperature. In one embodiment, the system is a closed system.In one embodiment, the system is an open system.

In one embodiment, the method increases the volume and/or weightcapacity of the storage and/or transport system by at least 1%, at least5%, least 10%, at least 15%, at least 20%, at least 25%, at least 50%,at least 60%, at least 70%, at least 80%, at least 90%, at least 100%,at least 150%, at least 200%, at least 300%, at least 400%, at least500%, at least 1000%, etc, relative to the standard values in the art.In one embodiment, the method allows for at least 1%, at least 5%, atleast 10%, at least 15%, at least 20%, at least 25%, at least 50%, atleast 60%, at least 70%, at least 80%, at least 90%, or 100% of thevolume of the system to be used for product storage.

In one embodiment, the method allows for a reduction in the energyrequired to store and/or transport the product of at least 1%, at least5%, at least 10%, at least 15%, at least 20%, at least 25%, at least50%, at least 60%, at least 70%, at least 80%, at least 90%, or 100%,relative to the standard values in the art. In one embodiment, themethod allows for a reduction in the refrigeration costs duringtransport of at least 1%, at least 5%, at least 10%, at least 15%, atleast 20%, at least 25%, at least 50%, at least 60%, at least 70%, atleast 80%, at least 90%, or at least 95%, relative to the standardvalues in the art. In one embodiment, the method allows for a reductionin the humidification/dehumidification costs during storage and/ortransport of at least 1%, at least 5%, at least 10%, at least 15%, atleast 20%, at least 25%, at least 50%, at least 60%, at least 70%, atleast 80%, at least 90%, or at least 95%, relative to the standardvalues in the art.

In one embodiment, the method allows for a reduction in the packagingcosts during storage and/or transport of at least 1%, at least 5%, atleast 10%, at least 15%, at least 20%, at least 25%, at least 50%, atleast 60%, at least 70%, at least 80%, at least 90%, or at least 95%,relative to the standard values in the art.

In one embodiment, the method allows for a reduction in the air, CO₂,N₂, hydrogen sulfide, steam, and other gas costs during storage and/ortransport of at least 1%, at least 5%, at least 10%, at least 15%, atleast 20%, at least 25%, at least 50%, at least 60%, at least 70%, atleast 80%, at least 90%, or at least 95%, relative to the standardvalues in the art.

In one embodiment, the standard capacity of a truck or container thattransports spinach is ⅓ full. In one embodiment, the method allows thetruck or container to be at least ⅔ full, etc., or 100% full of spinach.In one embodiment, the standard for a truck or container that transports“baby leaf” produce (spinach, kale, etc) is less than about 80%capacity. In one embodiment, the standard for truck or containercapacity for Broccoli is about 40-60% capacity (50% broccoli, remainderice slurry); Green onions are shipped at about 25-50% green onion,remainder ice; and Cilantro is shipped at 20-40% cilantro, remainderice. In one embodiment, the method allows for a reduction in the amountof ice. In one embodiment, the disclosure provides a method that allowstruck or container capacity (e.g., volume utilization) to increase by atleast 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least10%, at least 15%, at least 20%, at least 25%, at least 50%, at least60%, at least 70%, at least 80%, at least 90%, at least 95%, at least100%, at least 200%, at least 300% and percentages in between, relativeto standard capacity. In one embodiment, the method of the disclosureallows for transport without ice or with reduced amounts of ice. In oneembodiment, the disclosure provides a method that allows a reduction inthe amount of ice required for transport by at least 1%, at least 2%, atleast 3%, at least 4%, at least 5%, at least 10%, at least 15%, at least20%, at least 25%, at least 50%, at least 60%, at least 70%, at least80%, at least 90%, at least 95%, at least 100%, at least 200%, at least300% and percentages in between, relative to standard ice levels. In oneembodiment, the disclosure provides a method of improving transport bytrain, air, and/or sea.

In one embodiment, the produce is a plant. In one embodiment, the plantis a vegetable or herb. In one embodiment, the vegetable or herb isselected from the group consisting of arugula, asparagus, basil, beet,broccoli, brussel sprout, cabbage, carrot, cauliflower, celery, chard,chicory, chive, coriander, corn, dill, endive, garlic, kale, lavender,leek, lettuce, mint, mushrooms, oregano, parsley, parsnip, pea, peanuts,rosemary, spinach, soybean, squash, thyme, turnip, and yam. Examples ofother plants are provided elsewhere in the specification.

In one embodiment, the produce is a fruit. In one embodiment, the fruitis selected from the group consisting of alfalfa, apples, apricots,avocados, barley, beans, berries, blueberries, bananas, beans, citrus,cucumbers, damsons, eggplants, grapes, grapefruits, figs, kiwis,mangoes, melons, nectarines, oranges, papayas, peaches, pears, peppers,pineapples, plums, pumpkins, raspberries, rice, rye, sorghum,strawberries, sunflowers, tomatoes, wheat, and zucchinis, etc. Examplesof other fruits are provided elsewhere in the specification.

In one embodiment, the flower is selected from roses, carnations,orchids, tulips, narcissus, daffodils, anthurium, mimosa, gladiolus,lilies, ferns, freesia. In one embodiment, the flower is selected fromArabidopsis thaliana, Afrimay violet, alstromeria, anemone, aster,azalea, begonia, bellflower, bougainvillea, buttercup, cactus, camellia,carnation, chrysanthemum, clematis, cockscomb, columbine, cosmos,cyclamen, daffodil, dahlia, daisy, false cypress, forsythia, freesia,gardenia, gladioli, hibiscus, hollyhock, hydrangea, iris, lilac, lily,mum, peony, pelargonium, petunia, poinsettia, poppy, rose, saintpaulia,snapdragon, statice, sunflower, tulip, orchid, waxflower, and zinnia.Examples of other flowers are provided elsewhere in the specification.

In one embodiment, the method comprises contacting the surface of theproduct with silk fibroin, chitin, mono- and di-acyl glycerides and/orothers (edible) coatings prior to and/or during transport. In oneembodiment, at least some of the surface of the product is coated withsilk fibroin and/or any other (edible) coating. In one embodiment, atleast at least 1%, at least 5%, at least 10%, at least 15%, at least20%, at least 25%, at least 50%, at least 60%, at least 70%, at least80%, at least 90%, at least 95%, or 100% of the surface is coated withsilk fibroin and/or any other (edible) coating. In one embodiment, atleast 1%, at least 5%, at least 10%, at least 15%, at least 20%, atleast 25%, at least 50%, at least 60%, at least 70%, at least 80%, atleast 90%, at least 95%, or 100% of the surface is coated with one ormore mono- or diacyl glycerides.

In one embodiment, the coating decreases the temperature of the coatedproduct. In one embodiment, the coating decreases the respiration rateof the coated product. In one embodiment, the temperature and/orrespiration rate is decreased by at least 1%, at least 5%, at least 10%,at least 15%, at least 20%, at least 25%, at least 50%, at least 60%, atleast 70%, at least 80%, at least 90%, or at least 95% relative to thatof the uncoated product. Accordingly, the disclosure provides a methodof decreasing the temperature and/or respiration rate of a productcomprising coating it with silk protein and/or any other (edible)coating. In one embodiment, the method of decreasing the temperatureand/or respiration rate of the product comprises contacting the surfaceof the product with silk fibroin, chitin, mono- and di-acyl glyceridesand/or other (edible) coatings prior to and/or during transport. Inanother embodiment, the method is carried out to minimize water loss inthe harvested plant part. Water loss or transpiration, refers to watervapor movement from the harvested plant part to the environment.

In one embodiment, the silk fibroin, chitin, mono- and di-acylglycerides and/or other coatings (including edible coatings) provides aninsulative effect. For example, the insulative effect may allow thecoated product to reduce internal temperature increases compared to anuncoated product. In one embodiment, the insulative effect is increasedby at least 1%, at least 5%, at least 10%, at least 15%, at least 20%,at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, atleast 90%, or at least 95% relative to that of the uncoated product.Accordingly, the disclosure provides a method of improving insulation ofa product comprising coating it with silk fibroin, chitin, mono- anddi-acyl glycerides and/or other (edible) coatings.

In one embodiment, the silk coating and/or other (edible) coatingprovides a UV filtering effect. In one embodiment, the UV filteringeffect is increased by at least 1%, at least 5%, at least 10%, at least15%, at least 20%, at least 25%, at least 50%, at least 60%, at least70%, at least 80%, at least 90%, or at least 95% relative to that of theuncoated product. Accordingly, the disclosure provides a method ofimproving UV filtration of a product comprising coating it with silkfibroin, chitin, mono- and di-acyl glycerides and/or other (edible)coatings.

In one embodiment, the surface is coated prior to harvesting. In oneembodiment, the surface is coated during harvesting by the harvester. Inone embodiment, the surface is coated within at least 5 minutes, atleast 10 minutes, at least one hour, at least 12 hours, at least 48hours, or at least 72 hours from harvesting (i.e., prior to or after).In one embodiment, the surface is coated after harvest but prior totransport. In one embodiment, the surface is coated within 1 hour, 2hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10hours, 11 hours, 12 hours, 24 hours, 48 hours, or 72 hours fromtransport. In one embodiment, the surface is coated prior to, during,and/or after transport.

In one embodiment, a metric of the coated item selected from one or moreof visual inspection (e.g., yellowing, wilting, rot, color), structuralintegrity, microbial load (e.g., microbial growth), bounce back (e.g.,the ability of certain products (e.g., leafy greens) to recover from thenegative effects of wilting once washed through rehydration, which showsassists in showing whether the product was damaged) respiration rate,taste, temperature, insulative effect, UV filtering effect, and/or odor.In one embodiment, the yellowing of a coated item decreases compared toan uncoated item by at least 1%, at least 2%, at least 5%, at least 10%,at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, atleast 50%, at least 75%. In one embodiment, the wilting of a coated itemdecreases compared to an uncoated item by at least 1%, at least 2%, atleast 5%, at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 40%, at least 50%, at least 75%. In one embodiment,the rot of a coated item decreases compared to an uncoated item by atleast 1%, at least 2%, at least 5%, at least 10%, at least 15%, at least20%, at least 25%, at least 30%, at least 40%, at least 50%, at least75%.

In one embodiment, the coating can extend the shelf life of products instorage or a similar environment. In some embodiments, the coatings ofthe present technology can extend the shelf life of a product comparedto an uncoated product by, at least 2 percent, at least 3 percent, atleast 4 percent, at least 5 percent, at least 10 percent, and least 20percent, at least 30 percent, at least 50%, at least 75%, and at least100 percent, or at least any number in between. In one embodiment, thecoatings extend the shelf life by 2-3 times. In a further embodiment thecoatings may permit optimization of product processing by extending theshelf life of products. For example, a product could be harvested,coated, and stored to await similar product that is harvested later topermit efficient processing. This could reduce processing times,manpower needed for processing, cost for processing, and other similarefficiency gains.

In one embodiment, the coating may permit higher storage and/ortransportation temperatures for a product. For example, if a product isnormally refrigerated at about 40 degrees Fahrenheit it could be kept ata temperature from about 40-70 degrees Fahrenheit. In a furtherembodiment, the coating could enable a product to withstand short heatspikes, for example up to 100 degrees Fahrenheit for a product normallystored at about 40 degrees Fahrenheit. The coating would enable areduction in refrigeration costs. The coating would also enable productto handle unpredictable transportation and/or storage environments moreeasily.

In one embodiment, the silk fibroin, chitin, mono- and di-acylglycerides and/or other (edible) coatings are added to the producteither as a powder or as a solution in which powdered silk fibroin,chitin, mono- and di-acyl glycerides, and other (edible) coatings havebeen reconstituted within a solvent.

BRIEF DESCRIPTION OF THE FIGURES

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawings will be provided by the U.S. Patent and Trademark Officeupon request and payment of the necessary fee.

FIG. 1 shows wilting severity as a method for classifying the propertiesof the tested product.

FIG. 2 shows yellowing as a method for classifying the properties of thetested product.

FIG. 3 shows rot as a method for classifying the properties of thetested product.

FIG. 4 shows a diagram of different layers of a leafy green in a toteand how they are evaluated.

FIG. 5 shows product in a transit study on Day 1.

FIG. 6 shows product in a transit study on Day 6, with the top layerexposed, (Silk Coated represents a product treated with silk fibroin).

FIG. 7 shows product in a transit study on Day 6, with the middle of thetote layer exposed (Silk Coated represents a product treated with silk).

FIG. 8 shows a quantification of percentage of leaves with a yellowingscore of 1 or 2 for different treatment methods.

FIG. 9 shows a quantification of the percentage of leaves with a wiltingscore of 1 or 2 for different treatment methods.

FIG. 10 shows a quantification of the percentage of layers with a rotscore of 1 for different treatment methods.

FIG. 11 shows the different groups of kale on day 1 of a shelf lifestudy.

FIG. 12 shows the different groups of kale on day 9 of a shelf lifestudy.

FIG. 13 shows the different groups of kale on day 28 of a shelf lifestudy.

FIG. 14 shows a quantification of the yellowing scores for the leaves inthe shelf life study over time.

FIG. 15 shows a quantification of the yellowing scores for the leaves inthe shelf life study over time.

FIG. 16 shows a quantification of the yellowing scores for the leaves inthe shelf life study over time.

FIG. 17 shows a respiration testing setup used to collect respirationdata.

FIG. 18 shows a second respiration testing setup used to collectrespiration data.

DETAILED DESCRIPTION

The disclosure relates to methods and compositions for improvingtransportation of food, flowers, plants or the like.

Definitions

In order for the present disclosure to be more readily understood,certain terms are first defined below. Additional definitions for thefollowing terms and other terms are set forth throughout theSpecification.

As used in this Specification and the appended claims, the singularforms “a,” “an” and “the” include plural referents unless the contextclearly dictates otherwise.

Unless specifically stated or obvious from context, as used herein, theterm “or” is understood to be inclusive and covers both “or” and “and”.

The term “and/or” where used herein is to be taken as specificdisclosure of each of the two specified features or components with orwithout the other. Thus, the term “and/or” as used in a phrase such as“A and/or B” herein is intended to include A and B; A or B; A (alone);and B (alone). Likewise, the term “and/or” as used in a phrase such as“A, B, and/or C” is intended to encompass each of the followingembodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; Aand B; B and C; A (alone); B (alone); and C (alone).

The terms “e.g.,” and “i.e.” as used herein, are used merely by way ofexample, without limitation intended, and should not be construed asreferring only those items explicitly enumerated in the specification.

The term “edible coating” refers to a thin layer of edible material,which is formed as a protective coating on foods and may be consumedtogether with those products. These layers are applied in liquid formonto the food surface, usually by immersing the product in afilm-forming solution formed by the structural matrix. Edible andnon-edible coatings of the disclosure may provide a barrier to moisture,oxygen, carbon dioxide, and solute movement from the product. They notonly reduce the loss of moisture, they may also delay the process ofripening, and prevent microbial spoilage. Some (edible) coatings may becomposed of polysaccharides, proteins, lipids, or a combination of thesecompounds. Most often, compounds of each of the three categories need tobe combined. Materials may be added to improve the structural,mechanical, or handling properties, as well as to improve quality,flavour, colour, or nutritional properties of the coated product. Thesemay include plasticizers (increase the strength and flexibility of thecoating but may also increase the coating permeability to water vapourand gases. Examples of plasticizers include polyols (e.g., glycerol,sorbitol, mannitol, propylene glycol, and polyethylene glycol), sucrose,sucrose fatty acid esters, and acetylated monoglycerides.

The terms “or more”, “at least”, “more than”, and the like, e.g., “atleast one” are understood to include but not be limited to at least 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 20, 21,22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39,40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57,58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108,109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122,123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136,137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149 or 150,200, 300, 400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000 ormore than the stated value. Also included is any greater number orfraction in between.

Conversely, the term “no more than” includes each value less than thestated value. In one embodiment, “no more than 100” includes 100, 99,98, 97, 96, 95, 94, 93, 92, 91, 90, 89, 88, 87, 86, 85, 84, 83, 82, 81,80, 79, 78, 77, 76, 75, 74, 73, 72, 71, 70, 69, 68, 67, 66, 65, 64, 63,62, 61, 60, 59, 58, 57, 56, 55, 54, 53, 52, 51, 50, 49, 48, 47, 46, 45,44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27,26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9,8, 7, 6, 5, 4, 3, 2, 1, and 0. Also included is any lesser number orfraction in between.

The terms “plurality”, “at least two”, “two or more”, “at least second”,and the like, are understood to include but not limited to at least 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58,59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76,77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94,95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109,110,111, 112, 113, 114,115, 116, 117, 118,119, 120, 121, 122, 123, 124,125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138,139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149 or 150, 200, 300,400, 500, 600, 700, 800, 900, 1000, 2000, 3000, 4000, 5000 or more. Alsoincluded is any greater number or fraction in between.

Throughout the specification the word “comprising,” or variations suchas “comprises” or “comprising,” will be understood to imply theinclusion of a stated element, integer or step, or group of elements,integers or steps, but not the exclusion of any other element, integeror step, or group of elements, integers or steps. It is understood thatwherever embodiments are described herein with the language“comprising,” otherwise analogous embodiments described in terms of“consisting of” and/or “consisting essentially of” are also provided.The term “consisting of” excludes any element, step, or ingredient notspecified in the claim. In one embodiment, “consisting of” is defined as“closing the claim to the inclusion of materials other than thoserecited except for impurities ordinarily associated therewith.” A claimwhich depends from a claim which “consists of” the recited elements orsteps may not add an element or step. The terms “consisting essentiallyof” or “consists essentially” likewise has the meaning ascribed in U.S.Patent law and the term is open-ended, allowing for the presence of morethan that which is recited so long as basic or novel characteristics ofthat which is recited is not changed by the presence of more than thatwhich is recited, but excludes prior art embodiments.

Unless specifically stated or evident from context, as used herein, theterm “about” refers to a value or composition that is within anacceptable error range for the particular value or composition asdetermined by one of ordinary skill in the art, which will depend inpart on how the value or composition is measured or determined, i.e.,the limitations of the measurement system. In one embodiment, “about” or“approximately” may mean within one or more than one standard deviationper the practice in the art. “About” or “approximately” may mean a rangeof up to 10% (i.e., ±10%). Thus, “about” may be understood to be within10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, 0.01%, or0.001% greater or less than the stated value. In one embodiment, about 5mg may include any amount between 4.5 mg and 5.5 mg. Furthermore,particularly with respect to biological systems or processes, the termsmay mean up to an order of magnitude or up to 5-fold of a value. Whenparticular values or compositions are provided in the instantdisclosure, unless otherwise stated, the meaning of “about” or“approximately” should be assumed to be within an acceptable error rangefor that particular value or composition.

As described herein, the term “plant” refers to any living organismbelonging to the kingdom Plantae, including, but not limited to, trees,herbs, bushes, grasses, and vines. The term refers to both monocots anddicots. Exemplary plants include, but are not limited to, corn,potatoes, roses, apple trees, sunflowers, wheat, rice, bananas,tomatoes, pumpkins, squash, lettuce, cabbage, oak trees, guzmania,geraniums, hibiscus, clematis, poinsettias, sugarmaye, taro, duck weed,pine trees, Kentucky blue grass, zoysia, coconut trees, brassica leafyvegetables (e.g., broccoli, broccoli raab, Brussels sprouts, cabbage,Chinese cabbage (e.g., Bok Choy and Napa), cauliflower, cavalo,collards, kale, kohlrabi, mustard greens, rape greens, and otherbrassica leafy vegetable crops), bulb vegetables (e.g., garlic, leek,onion (dry bulb, green, and Welch), shallot, and other bulb vegetablecrops), citrus fruits (e.g., grapefruit, lemon, lime, orange, tangerine,citrus hybrids, pummelo, and other citrus fruit crops), cucurbitvegetables (e.g., cucumber, citron melon, edible gourds, gherkin,muskmelons (including hybrids and/or cultivars of cucumis melons),water-melon, maytaloupe, and other cucurbit vegetable crops), fruitingvegetables (including eggplant, ground cherry, pepino, pepper, tomato,tomatillo, and other fruiting vegetable crops), grape, leafy vegetables(e.g., romaine), root/tuber and corm vegetables (e.g., potato), and treenuts (e.g., almond, pemay, pistachio, and walnut), berries (e.g.,tomatoes, barberries, currants, elderberryies, gooseberries,honeysuckles, mayapples, nannyberries, Oregon-grapes, see-buckthorns,hackberries, bearberries, lingonberries, strawberries, sea grapes,lackberries, cloudberries, loganberries, raspberries, salmonberries,thimbleberries, and wineberries), cereal crops (e.g., corn, rice, wheat,barley, sorghum, millets, oats, ryes, triticales, buckwheats, fonio, andquinoa), pome fruit (e.g., apples, pears), stone fruits (e.g., coffees,jujubes, mangos, olives, coconuts, oil palms, pistachios, almonds,apricots, cherries, damsons, nectarines, peaches and plums), vines(e.g., table grapes and wine grapes), fibber crops (e.g. hemp andcotton), ornamentals, and the like. In some embodiments, the plant is anedible plant, or an edible plant part. In accordance with thisembodiment, the edible plant part is selected from the group consistingof alfalfa, apple, apricot, argula, asparagus, avocado, banana,blueberry, barley, basil, bean, beet, berries, blueberries, broccoli,brussel sprout, cabbage, carrot, cauliflower, celery, chard, chicory,chives, citrus, corn, coriander, cucumber, damson, dill, eggplant,endive, figs, garlic, grape, grapefruit, kale, kiwi, lavender, leek,lettuce, mango, mayola, melon, mint, mushroom, nectarine, oregano,orange, onion, papaya, parsley, parsnip, pea, peach, peanut, pear,pepper, pineapple, plum, potato, pumpkin, radish, raspberry, rice,rosemary, rye, sweet potato, sorghum, soybean, spinach, squash,strawberry, squash, sunflower, thyme, turnip, tomato, wheat, yam, andzucchini. The plant part may be selected from the group consisting of aflower, a fruit, a vegetable, and a herb.

Further, as used in the following, the terms “preferably”, “morepreferably”, “most preferably”, “particularly”, “more particularly”,“specifically”, “more specifically” or similar terms are used inconjunction with optional features, without restricting furtherpossibilities. Thus, features introduced by these terms are optionalfeatures and are not intended to restrict the scope of the claims in anyway. The disclosure may, as the skilled person will recognize, beperformed by using alternative features. Similarly, features introducedby “in an embodiment of the disclosure” or similar expressions areintended to be optional features, without any restriction regardingfurther embodiments of the disclosure, without any restrictionsregarding the scope of the disclosure and without any restrictionregarding the possibility of combining the features introduced in suchway with other optional or non-optional features of the disclosure.

The term “shelf life” means the duration of time that a product may bestored without becoming unfit for use, consumption, or sale. Shelf lifeis therefore the maximum time for which a product may be stored underexpected or specified conditions. The term “extending the shelf life”means that the coating prolongs the shelf life compared to a productwithout the coating when both products are processed and stored underidentical or substantially identical conditions. In some embodiments,the coatings of the present technology can extend the shelf life of aproduct compared to an uncoated product by, at least 2 percent, at least3 percent, at least 4 percent, at least 5 percent, at least 10 percent,and least 20 percent, at least 30 percent, and at least 100 percent, orat least any number in between. In one embodiment, the coatings extendthe shelf life by 2-3 times.

As described herein, any concentration range, percentage range, ratiorange or integer range is to be understood to be inclusive of the valueof any integer within the recited range and, when appropriate, fractionsthereof (such as one-tenth and one-hundredth of an integer), unlessotherwise indicated.

Units, prefixes, and symbols used herein are provided using theirSystème International de Unites (SI) accepted form. Numeric ranges areinclusive of the numbers defining the range.

Description

Produce grower/shippers and processors can see upwards of 30% productlosses throughout the supply chain, even despite a highly efficient coldchain. Some companies ship leafy greens (spinach, arugula, baby kale,etc.) from their growing regions to their processing facilities at lowdensity to minimize spoilage. This results in low freight capacityutilization. This disclosure provides means to mitigate this issuethrough natural and edible food coatings that extend shelf life bycombating the major mechanisms of spoilage: microbial growth, oxidation,moisture loss, and/or gas exchange. In one embodiment, the disclosureimplements a solution post-processing and/or prior to packing. In oneembodiment, the disclosure provides the impact of Cambridge Crops'products (e.g., silk fibroin coating) in driving supply chainefficiencies: reduced shrink; increased freight capacity utilization,reducing the number of trucks on the road, reductions in cost and carbonemissions and other environmental advantages. In one embodiment, thecoating is applied immediately post-harvest (e.g., by the harvester(e.g., any human or machine that can assist in harvesting productincluding but not limited to a harvester, combine, threshers, strippercleaners, stripper loaders, tree shakers, picking machines,spinners/digger, huller, husker, conveyors, augers, reaper-binders,swathers, etc.) during a wash cycle). In one embodiment, the coating isapplied pre-harvest. In one embodiment, the coating is applied after theproduce has been washed. In one embodiment, the coating is appliedduring the wash process (e.g., during the third wash in a triple washcycle). In one embodiment, the coating is applied prior to placing theproduct in packaging. In one embodiment, the coating is applied afterthe product is shipped to a distribution center.

In one embodiment, the transport totes are packed with greens to 100%utilization. In one embodiment, the greens are exposed to short heatspikes to simulate inhomogenous cooling in field as well as cold-chainbreaks over the course of 6 days.

In one embodiment, the metrics are selected from one or more of visualinspection (e.g., yellowing, wilting, rot, color), structural integrity,microbial load (e.g., microbial growth), bounce back (e.g., the abilityof certain products (e.g., leafy greens) to recover from the negativeeffects of wilting once washed through rehydration, which shows assistsin showing whether the product was damaged), respiration rate, taste,temperature, insulative effect, UV filtering effect, and/or odor. In oneembodiment, the disclosure provides a method to provide for alternativeshipment methods that may be longer than traditional shipment methods.The alternative methods are made possible by the use of the coating,which extends the shelf life and/or limits the decomposition of theproduct.

Accordingly, in one embodiment, the products are contacted with one ormore coating agents. In one embodiment, the coating agents are combinedwith pulsed light treatments, high hydrostatic pressure,gamma-radiation, ozone, UV light, modified atmospheric packaging, whichmay work additively or synergistically with the coating agents. In oneembodiment, the coating agents are edible. Edible coatings and otherplant coatings may be fabricated from different materials, which may begenerally classified into three categories: hydrocolloids, includingpolysaccharides and proteins; lipids; and composite materials,consisting of a combination of different hydrocolloids or hydrocolloidsand lipids, to take advantage of the complementary functional propertiesof the different constitutive materials or overcome the respectivedrawbacks.

In one embodiment, the product is contacted (e.g., coated) with silkprotein. In one embodiment, the product is contacted (e.g., coated,mixed with) with one or more of the following coating agents to improvestorage and/or transport: high fructose corn syrup, starch, acetylatedglycerides (e.g., mono- and/or di-acyl glyceride(s)), a mixture ofmonoacylglycerides (i.e., monoglycerides or fatty acid monoesters ofglycerol), primarily 2,3-dihydroxypropyl palmitate,1,3-dihydroxypropan-2-yl palmitates. In one embodiment, the coating isselected from one or more of lipids, resins, polysaccharides, proteins(e.g., silk, soy, whey, rice bran extract, egg albumin and wheatprotein), polymers, composites, bilayer compositions, plastisizers(e.g., low molecular eight polyols), antifoam agents (which may beselected from alkyl poly acrylates, fatty acids, fatty alcohols,monoglycerides, diglycerides, triglycerides, a silicone-based foamcontrol agent, and mixtures thereof), surfactants (e.g., lecithin andlecithin derivatives, acetylated monoglycerides (e.g., mono- and/ordi-acyl glyceride(s)), ethylene glycol, monostearate, glycerolmonostearate, and sorbitan fatty acid esters (Tweens)), and emulsifiers.The term “silicone-based foam control agent” refers to a polymer with asilicon backbone. In one embodiment, the foam control agent is asilicone-based foam control agent. Suitable silicone-based foam controlagents include, but are not limited to, polydimethylsiloxane fluid andpolydimethylsiloxane-treated silica. In one embodiment, the coating is amixture of one or more film-forming agent with a low molecular weightmolecule. As used herein, the term “glyceride” refers to esters whereone, two, or three of the —OH groups of the glycerol have beenesterified. Monoglycerides, diglycerides, and triglycerides may compriseesters of any of the fatty acids described herein. Examples of suitablefatty acids are saturated or unsaturated and may be obtained fromnatural sources (e.g., palm oil, coconut oil, babassu oil, saffloweroil, tall oil, castor oil, tallow and fish oils, grease, and mixturesthereof) or may be synthetically prepared. Examples of suitable fattyacids for use in the present invention include capric acid, lauric acid,myristic acid, palmitic acid, stearic acid, arachidic acid, and behenicacid. In one embodiment, the coating includes a mixture of any of thelisted agents.

In one embodiment, the carbohydrate materials are selected fromcellulose, starch, pectin, alginate, carragean, furcellaran, chitosam,gum Arabic, gum ghatti, gun karaya, guar gum, locust bean gum, xanthangum, gellan gum, gum tragamayth, wax (beeswax, paraffin and polyethylenewax, and other petroleum-based products), resin (shellac, wood rosin,coumarone). In one embodiment, the protein material is selected fromsilk protein, soy protein, zein (corn), casein, whey, wheat gluten, ricebran extract, egg albumin, and peanut protein.

In one embodiment, the (edible) coating is selected from Aloe vera gel,basil seed mucilage, gum Arabic, fungal chitosan, gelating, chitosan,guar gum, sodium alginate, calcium chloride, xanthan gum,carboxymethylcellulose, stearic acid, chitosan hydrochloride, cassavastartch (which may be reinforced by startch nanocrystals), Aloearborescens gel, fucoidan, maltodextrins, whey protein isolates,nanocellulose, pectin, corn flour, ethanolic extracts of the leaves andpods of cocoa, leaves, and hulls of coffee, locust bean gum, sesameproteins, whey protein nanofibrils, sweet potato starch,transglutaminase, walnut flour protein, and mixtures thereof. In oneembodiment, one or more of these compounds is combined with silk fibroinprotein.

In one embodiment, the coating impacts one or more of the following:delay ripening of the product (e.g., fruit, vegetable), delay colorchange, reduce water loss, reduce decay, improve appearance, reducechilling and/or mechanical injury, and add shine or gloss. In oneembodiment, ripening is a process that includes changes in color,flavor, and texture (e.g. softening). In one embodiment, the coatingscarry natural antagonists to pathogens (e.g., nature seal (NS), NS+US 7,NS+Imazali, shellac+Imazalie). In one embodiment, the disclosureprovides coatings that may be used as carriers of other usefulingredients such as color or aroma additives, antimicrobials, flavours(e.g., vanilla essence), nutraceuticals and anti-oxidants. Ingredientssuch as a nutraceutical agent or an immune response enhancer may also beadded to provide additional properties to the coating. The nutraceuticalagent typically comprises at least one probiotic, examples of whichinclude Lactobacillus acidophilus, Lactobacillus casei and,Bifidobacterium lactic. An example of immune response enhancer includesa yeast gluco polysaccharide. In some embodiments, these additionalingredients are incorporated into the coating solution to furtherimprove the quality, stability, and safety of the product.

In one embodiment, the product is climacteric fruit. In one embodiment,the product is non-climacteric fruits. Climacteric fruit continue toripen after harvest, whereas non-climacteric do not. Climateric producetypically requires expedited shipment.

In one embodiment, the product is further exposed to temperaturecontrol. In one embodiment, temperature control is used because it mayaffect the rate of fruit respiration. In general, higher temperaturesincrease, and lower temperatures decrease fruit and vegetablerespiration rates.

In one embodiment, the product is exposed to controlled atmospherestorage and/or modified atmosphere packaging. In one embodiment, thesemethods are combined with one or more of the coatings of the disclosure.

In one embodiment, the silk compositions of the present disclosurecomprise natural and edible silk coatings that may extend the shelf lifeof perishable goods and decrease reliance on energy and cost intensivecold chains. The silk coatings of the present disclosure may beodorless, low cost, edible, compostable, and biodegradable. Silk fibroinis a structural protein that may be produced and extracted fromsilkworm, spiders, or other insects. It may also be otherwise generatedsynthetically. Silk fibroin is naturally produced by species such as,without limitation, Antheraea mylitta; Araneus bicentenarius; Araneusventricosus; Bombyx mori; Bombyx mandarins; Galleria mellonella; Nephilaclavipes; Nephila madagascariensis; and Tetragnatha versicolor. Silkfibroin's unique properties are derived from its structure, consistingof hydrophobic blocks separated by hydrophilic spacers. In its naturalstate, silk fibroin is organized in beta-sheets, which are formed byhighly ordered crystalline regions alternated by amorphous regions. Thisunique structure results in high levels of strength and toughness forsilk fibroin-based materials. The wide range of forms into which silkfibroin solution may be processed make it attractive for severalhigh-tech applications, including scaffolds for tissue engineering, bonescrews for fixation, and drug depots for therapeutic delivery. In someembodiments, the silk compositions are as described in U.S. PatentApplication Publication No. US 2020-0178576 A1 and U.S. patentapplication Ser. No. 17/230,822, incorporated herein by reference intheir entirety. The silk fibroin may be isolated from silk sericin andthe protein coat through the Ajisawa method or through other methodsusing water and salts, including chaotropic salts. In some embodiments,silk fibroin may be prepared according to the method described inMarelli, B., Brenckle, M., Kaplan, D. et al. Silk Fibroin as EdibleCoating for Perishable Food Preservation. Sci Rep 6, 25263 (2016),https://doi.org/10.1038/srep25263, incorporated herein by reference inits entirety.

In one embodiment, the silk fibroin may be a dried powder. In somefurther embodiments, the silk fibroin may be in liquid form. In someembodiments, the solution may comprise the silk fibroin powder mixedwith a solvent. In some embodiments, the solvent may be a liquid. Insome further embodiments, the solvent may be an acid with a pH under6.9. Alternatively, the solvent may be an alcohol or water. In otherembodiments, the solvent may be acetic acid. In some embodiments, thesilk fibroin powder may be in a mixture containing an additive. In somealternative embodiments, the liquid solvent may contain an additive. Insome embodiments, both the silk fibroin powder mixture and the liquidsolvent may contain additives. In some alternative embodiments, the silkfibroin may be emulsified with the additive prior to being mixed intothe solution. In one embodiment, liquid silk fibroin may be mixed ordry-blended with the additive prior to being mixed into the solution. Insome embodiments, the additive may be at least one of a sugar, aplasticizer, or a crosslinking agent. In one embodiment, the sugaradditive may be a sugar-ol, a poly-ol, or a hygroscopic polymer (e.g.,polyethylene glycol). In other examples, if the sugar additive is acrosslinking agent, the crosslinking agent may be photoreactive.Specifically, the crosslinking agent may be, for example, one or more ofhorseradish peroxidase, lysyl oxidase, disuccinimidyl suberate,disuccinimidyl glutarate, N-hydroxysuccinimide ester, or an aryl azide.In some alternative embodiments, the additive may include one or more ofa bacterium, a metal, an enzyme, or a biologic. In one embodiment, themetal may include one or more of an alkali metal, an alkaline earthmetal, or a transition metal. In one embodiment, the enzyme may includeerepsin maltase, lactase, sucrase, disaccharidases, lingual lipase,lysozymes, salivary amylase, pepsin, gastric lipase, other lipases,hydrochloric acids, intrinsic factors, mucins, gastrins, trypsinogen,ductal cells, carboxypeptidase, elastases, and the like. In some otheralternative embodiments, the additive may be at least one of a coloringagent, a chelator, a ligand, an antimicrobial, a filler, a scent, or aflavor. In one embodiment, the coloring agent may be allura red, Ponceau3R, amaranth, erythrosine, indigotine, Light Green SF, Naphthol yellow,Orange 1, quinoline yellow, tartrazine, an E1 suit (e.g., E100, E161b,etc.), an anthocyanin, a betacyanin, a carotenoid, or a phenolic. Inother examples, the chelator may be ethylenediaminetetraacetic acid(EDTA), transferrin, or desferrixoxamine. In other examples, themicrobial may be acetic acid, benzoic acid, natamycin, nisin, nitrate,nitrite, propionic acid, sorbic acid, sulfite, or sulfur dioxide. Inother examples, the filler may be cellulose. In other alternativeembodiments, the additive may be at least one of a vitamin, a nutrient,an antioxidant, and a protein. In one embodiment, the protein may be apeptide, an amino acid, (e.g., a post-translated amino acid), or asynthetic amino acid. A nutrient may be defined as a mineral, protein,carbohydrate, fat, Q10, glutathione, lithium, probiotic, glycine, DHA,flavonoid, or others. An antioxidant may include vitamins C and E,selenium, carotenoids, thiols, catalase, superoxide dismutase, uricacid, and ubiquinol. In some further alternative embodiments, theadditive may be at least one of a green tea extract, a rosemary extract,a phenolic antioxidant, catechin, acerola, tocopherol, chamomileextract, malphigia emarginata, Camellia sinensis, epicatechin,epigallocatechin, gallochatechin, epigallocatechin gallates, vitamin A,vitamin E, vitamin D, vitamin K, and/or vitamin C. In some embodiments,the additive may be mixed with an accelerant or an excipient. In oneembodiment, the additive may be mixed with polyethylene glycol orxylitol. In some further embodiments, the additive may be emulsifiedwith the accelerant or excipient and mixed into a silk fibroin solution.

In some further embodiments, the solution may be deposited onto theproduct via spray-coating. Alternatively, the solution may be depositedonto the product via dip-coating. In some embodiments, the silk isapplied to the product by submersion, coating (e.g., doctor blade,surface coating), washing, tablet coating, and/or lamination. In someother embodiments, the silk is applied to a product by spray coating,curtain coating, or slot coating. In some embodiments, the silk fibroinmay not be annealed after or before deposition. In some furtherembodiments, the product may include multiple layers of silk fibroin. Inone embodiment; the product could be sprayed with silk fibroin solution,dried, and then sprayed once more. This may happen any number of timesto add thickness and additional layers. In some further embodiments, theproduct may comprise multiple layers, with each layer serving afunction. In one embodiment, the product may be coated with silkfibroin. Then, the silk fibroin-coated product may be itself coated byanother coating that is hydrophobic or water-tight such that water maynot permeate the outer layer and reach the inner silk fibroin layer. Insome further embodiments, a tablet-coating may be utilized, where thesilk fibroin is coated while in an industrially-relevant drum. Tabletcoating may additionally be utilized, as well as film-coating. Acombination of the processes and methods disclosed above could be usedto obtain the desired coating performance on any suitable product.

Coatings may be produced by wet (e.g., evaporation of the solvent) ordry methods. Drying may be done by heat conduction or convention orinfrared heating. In one embodiment, the coating is applied by one ormore of dipping, immersing, spraying, spreading, and/or saturatedbrushes.

In one embodiment, of the above, the coating may be applied to thesurface of the product for between 1 and 3,600 seconds, for examplebetween 1 and 3000 seconds, between 1 and 2000 seconds, between 1 and1000 seconds, between 1 and 800 seconds, between 1 and 600 seconds,between 1 and 500 seconds, between 1 and 400 seconds, between 1 and 300seconds, between 1 and 250 seconds, between 1 and 200 seconds, between 1and 150 seconds, between 1 and 125 seconds, between 1 and 100 seconds,between 1 and 80 seconds, between 1 and 60 seconds, between 1 and 50seconds, between 1 and 40 seconds, between 1 and 30 seconds, between 1and 20 seconds, between 1 and 10 seconds, between 5 and 3000 seconds,between 5 and 2000 seconds, between 5 and 1000 seconds, between 5 and800 seconds, between 5 and 600 seconds, between 5 and 500 seconds,between 5 and 400 seconds, between 5 and 300 seconds, between 5 and 250seconds, between 5 and 200 seconds, between 5 and 150 seconds, between 5and 125 seconds, between 5 and 100 seconds, between 5 and 80 seconds,between 5 and 60 seconds, between 5 and 50 seconds, between 5 and 40seconds, between 5 and 30 seconds, between 5 and 20 seconds, between 5and 10 seconds, between 10 and 3000 seconds, between 10 and 2000seconds, between 10 and 1000 seconds, between 10 and 800 seconds,between 10 and 600 seconds, between 10 and 500 seconds, between 10 and400 seconds, between 10 and 300 seconds, between 10 and 250 seconds,between 10 and 200 seconds, between 10 and 150 seconds, between 10 and125 seconds, between 10 and 100 seconds, between 10 and 80 seconds,between 10 and 60 seconds, between 10 and 50 seconds, between 10 and 40seconds, between 10 and 30 seconds, between 10 and 20 seconds, between20 and 100 seconds, between 100 and 3,000 seconds or between 500 and2,000 seconds.

Properties of the coating, such as thickness, cross-link density, andpermeability, may be varied to be suitable for a particular product byadjusting the specific composition of the coating agent, the specificcomposition of the solvent, the concentration of the coating agent inthe solvent, and the conditions of the coating deposition process. Theconcentration of the solute (e.g., coating agent) in the solvent may,for example, be in a range of about 0.5 mg/mL to 200 mg/mL.

In one embodiment, the coating is applied to the product as a whole. Inone embodiment, the coating is applied to a part of the product. In oneembodiment, the coating is applied prior to harvest. In one embodiment,the coating is applied after and/or during harvest. In one embodiment,the product, or a part thereof, by be harvested within 1 hour, 2 hours,3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 48, 72 hours, orany time and time interval in between or after a coating is applied.

In one embodiment, the coating agent is prepared in an aqueous solution.In one embodiment, the coating agent is prepared in an alcohol solution.In one embodiment, the coating agent is prepared in an organic acidsolution. Examples of organic acids include citric acid, malic acid,tartaric acid, acetic acid, lactic acid, and levulinic acid. In oneembodiment, the coating solution may comprise 1.00 to 3.00 wt %, 1.00 to2.50 wt %, 1.00 to 2.00 wt %, 1.50 to 2.00 wt %, or 1.50 to 2.50 wt %coating agent (e.g, silk). In one embodiment, the coating solution maycomprise 0.05 to 0.10 wt %, 0.05 to 0.15 wt %, 0.05 to 0.50 wt %, 0.05to 0.15 wt %, 0.10 to 0.40 wt %, 0.20 to 0.30 wt % coating agent. In oneembodiment, the coating solution may comprise 0.05 to 0.10 wt %, 0.05 to0.15 wt %, 0.05 to 0.50 wt %, 0.05 to 0.15 wt %, 0.10 to 0.40 wt %, 0.15to 0.35 wt %, 0.20 to 0.30 wt % coating agent. In one embodiment, theaqueous solution may comprise from 90 to 99 wt %, 92 to 99 wt %, 94 to99 wt %, 96 to 99 wt %, 97 to 99 wt %, or 98 to 99 wt % of water orother solvent. In one embodiment, the coating solution may comprise1%-20%, or more preferably 2%-7%, or more preferably 2.5%-5% coatingagent (e.g., silk). In one embodiment, the agent is present at aconcentration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24.25, 26, 27, 28, 29, 30, 31, 32, 33.34,35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70,71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88,89, 90, 91, 92, 93, 94, 95, 96, 96, 97, 98, 99, 100, 200, 300, 400, 500,600, 700, 800, 900, 1000 micrograms/ml. In one embodiment, the agent ispresent at a concentration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39.40, 41, 42, 43, 44, 45, 46, 47, 48, 49,50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67,68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85,86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 96, 97, 98, 99, 100, 200,300, 400, 500, 600, 700, 800, 900, 1000 mg/ml. In one embodiment, theagent is present at a concentration of at least 1, at least 2, at least3, at least 4, at least 5, at least, at least 7, at least 8, at least 9,at least 10, at least 11, at least 12, at least 13, at least 14, atleast 15, at least 16, at least 17, at least 18, at least 19, at least20, at least 21, at least 22, at least 23, at least 24, at least 25, atleast 26, at least 27, at least 28, at least 29, at least 30, at least31, at least 32, at least 33, at least 34, at least 35, at least 36, atleast 37, at least 38, at least 39, at least 40, at least 41, at least42, at least 43, at least 44, at least 45, at least 46, at least 47, atleast 48, at least 49, at least 50, at least 51, at least 52, at least53, at least 54, at least 55, at least 56, at least 57, at least 58residues, at least 59, at least 60, at least 61, at least 62, at least63, at least 64, at least 65, at least 66, at least 67, at least 68, atleast 69, at least 70, at least 71, at least 72, at least 73, at least74, at least 75, at least 76, at least 77, at least 78, at least 79, atleast 80, at least 90, or 100% w/v, or w/w, in the coating solution.

Applying Silk Fibroin to a Product

Cocoons from the Bombyx mori may be used as source of silk fibroin. Thesilk fibroin may be extracted from the cocoons during a multi stepprocess. As a non limiting example of this process, the first step mayinclude degumming, by exposing the cocoons to a compound (e.g., sodaash) to separate the sericin from the cocoons. Next, the cocoons arerinsed with a fluid (e.g., water) to remove the sericin, resulting incocoons that are primarily made of silk fibroin. The cocoons are thenexposed to a second compound (e.g., a chaotropic agent) which dissolvesthe silk fibroin into a solution (e.g., water). The solution is thenpurified to remove the second compound (e.g., by dialysis, centrifuging,or other filtering method, including combinations of the above) to yielda solution made primarily of water and solubized silk fibroin. Thesolution is then sterilized, either by pasteurization ormicrofiltration, or both, to remove all aggregates, reduce turbidity,and kill microbes. The final concentration of the silk fibroinsuspension can be adjusted to include a specific wt %, for example about1%-20%, or more preferably 2%-7%, or more preferably 2.5%-5%. In someembodiments, the silk fibroin solution is then turned into powder by theuse of a spray dryer or other suitable device. Other suitable processesto create the silk fibroin solution could be used.

However, other concentrations of silk fibroin could be used, dependingon the food product and the implementation necessary. Similarly,different weight average molecular weight (M_(w)) silk fibroin solutionscould be utilized. In some embodiments, the fibroin fragments comprise aweight average molecular weight (M_(w)) from about 1 kDa to about 600kDa. In some embodiments, the molecular weight (MW) of the silk fibroinfragments may range from about 10 kDa to about 1000 kDa, withconcentrations of fragments of different molecular weight (MW).

A product can be coated using various different methods. The first stepis creating a solution that contains silk fibroin as described above. Insome embodiments, the silk fibroin solution is manufactured directly. Inother embodiments, the silk fibroin is turned into a powder, that isthen reconstituted into a solution (e.g., water) to create the silkfibroin solution.

Once the silk fibroin solution is created, the product can be coatedusing various different techniques depending on different variables,such as the type of product, the desired coating type, coverage,thickness, or number of layers, the shipping/storage conditions, theshipping/storage duration, etc. The silk solution may be applied to aproduct by any suitable method. In some embodiments, the silk is appliedto the product by submersion, coating (e.g., spray, airbrush, doctorblade, dip coating, surface coating), washing, tablet coating, and/orlamination. In some embodiments, the silk is applied to a product byspray coating, curtain coating, or slot coating. In some embodiments,the silk is deposited onto a product and then dried via, for example,air-drying, freeze-drying, vacuum drying, or heat-drying. In someaspects, the silk solution may be deposited onto a product viaspray-coating. Alternatively, the solution may be deposited onto aproduct via dip-coating. In some aspects, the silk may be applied byelectrospinning.

In some aspects, the product may undergo pre-treatment, by for examplebeing exposed to a basic solution and then dried prior to application ofthe silk fibroin solution. By a further example, a pre-treatment couldinclude adding a binder and/or adhesive to the product to improve theabrasion resistance or adhesion of the silk solution.

In some aspects, the silk fibroin coating applied to a product mayundergo post-treatment to improve the properties of the silk fibroincoating, including treatment with alcohol (e.g., ethanol, isopropanol,or other alcohols). In another aspect, the post-treatment could includeultraviolet exposure to cross-link the silk coating or heat treatment toactivate a foaming agent in the silk coating. In some aspects, the silkmay or may not be annealed after or before deposition, including bywater annealing.

In some further aspects, the product may include multiple layers ofsilk. In one embodiment, the product may be sprayed with a silk fibroinsolution, dried, and then sprayed once more. This may happen any numberof times to add thickness and additional layers. In some furtheraspects, the product may comprise multiple layers, with each layerserving a function. For example, the product may be coated with silksolution. Then, the silk solution coated product may be itself coated byanother coating. Such other coating may be hydrophobic. A combination ofthe processes and methods disclosed above could be used to obtain thedesired coating performance on any suitable product.

The silk coating may be of different thicknesses depending on thedesired application or method used to create it. In some aspects, thesilk coating may be uniform in thickness. In some aspects, the silkcoating may not have a uniform thickness. For example, the silk coatingmay be thinner in some areas due to natural variances caused by themethod used to apply the coating (e.g., a spray coating may vary inthickness due to variances during the application process such asspraying one area slightly longer than others resulting in differentcoating thickness). In other aspects, the thickness may be varied onpurpose to create specific barrier properties. In some embodiments thethickness may range from about 10 nm to about 1 mm, from about 10 nm toabout 50 nm, from about 50 nm to about 100 nm, from about 100 nm toabout 200 nm, from about 100 nm to about 100 μm, from about 200 nm toabout 300 nm, from about 300 nm to about 500 nm, from about 500 nm toabout 1 μm, from about 1 μm to about 5 μm, from about 5 μm to about 10μm, from about 10 μm to about 15 μm, from about 10 μm to about 20 μm,from about 10 μm to about 25 μm, from about 10 μm to about 50 μm, fromabout 10 μm to about 100 μm, from about 15 μm to about 25 μm, from about15 μm to about 35 μm, from about 15 μm to about 50 μm, from about 20 μmto about 40 μm, from about 20 μm to about 50 μm, from about 25 μm toabout 35 μm, from about 25 μm to about 50 μm, from about 25 μm to about100 μm, from about 30 μm to about 50 μm, from about 30 μm to about 75μm, from about 40 μm to about 50 μm, from about 40 μm to about 75 μm,from about 50 μm to about 65 μm, from about 50 μm to about 75 μm, fromabout 50 μm to about 85 μm, from about 50 μm to about 100 μm, from about100 μm to about 150 μm, from about 100 μm to about 200 μm, from about100 μm to about 250 μm, from about 150 μm to about 250 μm, from about100 μm to about 300 μm, from about 100 μm to about 400 μm, from about100 μm to about 500 μm, from about 200 μm to about 400 μm, from about200 μm to about 500 μm, from about 400 μm to about 600 μm, from about500 μm to about 800 μm from, from about 500 μm to about 1 mm, from about100 μm to about 1 mm.

In an embodiment, the thickness of the silk coating may have a thicknessless than 1 mm. In an embodiment, the thickness of the silk coating mayhave a thickness less than 500 μm. In an embodiment, the thickness ofthe silk coating may have a thickness less than 300 μm. In anembodiment, the thickness of the silk coating may have a thickness lessthan 200 μm. In an embodiment, the thickness of the silk coating mayhave a thickness less than 150 μm. In an embodiment, the thickness ofthe silk coating may have a thickness less than 100 μm. In anembodiment, the thickness of the silk coating may have a thickness lessthan 80 μm. In an embodiment, the thickness of the silk coating may havea thickness less than 70 μm. In an embodiment, the thickness of the silkcoating may have a thickness less than 60 μm. In an embodiment, thethickness of the silk coating may have a thickness less than 50 μm. Inan embodiment, the thickness of the silk coating may have a thicknessless than 40 μm. In an embodiment, the thickness of the silk coating mayhave a thickness less than 30 μm. In an embodiment, the thickness of thesilk coating may have a thickness less than 25 μm. In an embodiment, thethickness of the silk coating may have a thickness less than 20 μm. Inan embodiment, the thickness of the silk coating may have a thicknessless than 15 μm. In an embodiment, the thickness of the silk coating mayhave a thickness less than 10 μm. In an embodiment, the thickness of thesilk coating may have a thickness less than 1 μm. In an embodiment, thethickness of the silk coating may have a thickness less than 500 nm. Inan embodiment, the thickness of the silk coating may have a thicknessless than 100 nm.

In some aspects, a product may be coated in its entirety or onlypartially. For example, if the product has a flat shape, such as forkale or spinach, it may be coated on both sides. As a further example,if the product is piece of produce, such as broccoli, it may only becoated on the outside surfaces, with the inner surfaces receiving littleto no coating. In another aspect, the product may only be coated over aportion of a surface of a product, for example a surface may only bepartially coated such that half of one surface receives a coating. In anembodiment, the product may have about 100% of its total surface areacoated, or about 80% or more of its total surface area coated, or about60% or more of its total surface area coated, or about 50% or more ofits total surface area coated, or about 40% or more of its total surfacearea coated, or about 30% or more of its total surface area coated, orabout 20% or more of its total surface area coated, or about 10% or moreof its total surface area coated. In some aspects, between about 80% toabout 100% of the total surface area of a product may be coated, orbetween about 60% to about 80% of the total surface area of a productmay be coated, or between about 50% to about 80% of the total surfacearea of a product may be coated, or between about 40% to about 60% ofthe total surface area of a product may be coated, or between about 20%to about 40% of the total surface area of a product may be coated, orbetween about 1% to about 20% of the total surface area of a product maybe coated.

A combination of the processes and methods disclosed above could be usedto obtain the desired coating performance on any suitable product.

Additional embodiments may be derived from the additional attachments.

EXAMPLES Example 1 Stressed Transit Study

A model was developed to mimic stressed conditions for transport ofgreens (e.g., spinach, kale, lettuce, etc.) to test the performance ofsilk fibroin coatings against uncoated greens and/or greens coated withwater. Stressed conditions representing increased metabolicrate/premature spoilage were created by recreating increased freightutilization by packing the greens more densely, exposing greens to“extreme” supply chain conditions using heat spikes (e.g., increases intemperature beyond normal temperatures encountered during transit) andthen comparing control versus treated groups to test the effects of silkfibroin.

In one exemplary experiment regarding greens, the greens (kale) werepacked in totes to 100% utilization, as shown in FIG. 5 . In normalconditions the totes are packed to about 10-19 pounds, but in thisexperiment the totes were packed to over 21 pounds, representing anincrease in weight of about 13% to about 100%. In total, three samplegroups were prepared: a control sample group (kale not treated with anysubstance); a water sample group (kale treated with water); and a silkcoated sample group (kale treated with silk fibroin solution). The waterand silk fibroin were applied via spraying. After application, all threegroups were hand tossed to better distribute the coating on the waterand silk coated sample group and to ensure that all groups were handledin the same manner. The application of the coating and or thedistribution method could be changed to create different transitenvironments with different stresses.

The three sample groups were then exposed to a simulated transit testwhere they were stored at regular temperatures of about 40 degreesFahrenheit. The samples were then exposed to 3 heat spikes. Thetemperature of the heat spike was about 100 degrees Fahrenheit and itwas held steady for about 30 minutes. The heat spikes were introducedevery 24 hours after the start of the experiment (starting aftertreatment of the kale), so the first occurred at 24 hours, the secondoccurred at 48 hours, and the third occurred at 72 hours. The threesample groups were returned to the regular storage temperature aftereach heat spike, and until the conclusion of the study, which was sixdays after treatment. FIG. 6 shows the top layer of the totes of thedifferent groups on day 6. FIG. 7 shows the middle layer of the totes ofthe water sample group and the silk coated sample group on day 6. Themiddle layer is the portion of the container most susceptible to rot dueto the increased density of the greens. To better illustrate thedifferent layers of the totes. FIG. 4 shows the layers of a leafy greenwhen held in a tote, with a top, middle, and bottom layer identified.

At the conclusion of the experiment the greens were scored to determineif the groups differed. Multiple different aspects of the product, inthis case kale, can be observed and compared, for example, metricsselected from one or more of visual inspection (e.g., yellowing,wilting, rot, color), structural integrity, microbial load (e.g.,microbial growth), bounce back (e.g., the ability of certain products(e.g., leafy greens) to recover from the negative effects of wiltingonce washed through rehydration, which shows assists in showing whetherthe product was damaged), respiration rate, taste, temperature,insulative effect, UV filtering effect, and/or odor.

In this experiment wilting, yellowing, and rot were compared across thedifferent groups (e.g., control, water, silk solution). The groups werecompared by randomly selecting 20 leaves and assigning them a numberbased on the scoring scales shown in FIGS. 1-3 . FIG. 6 shows theresults for yellowing of the leaves after six days for the three groups.The control group on the far left faired the worst, followed by thewater group in the middle, with the silk fibroin solution group (SilkCoated) on the right performing the best. This improvement wasnoticeable at the middle layer of the totes too, as seen in FIG. 7 ,which shows the middle layer of the water (left) and silk fibroinsolution (right) groups after six days. This is illustrated more clearlyin FIG. 8 , which shows the percentage of leaves in the various groupswith a yellowing score of 1 or 2. A score of 1 or 2 was awarded if theleaf had yellowing on 0-10% of leaf surface or on 10-30% of the leafsurface, respectively, as judged in FIG. 2 . As FIG. 8 shows, about 93%of the leaves treated with silk fibroin coated achieved a score of 1 or2, while the control group and water group had approximately 69% and82%, respectively, of leaves with the same score. FIG. 9 shows theresults for wilting across the different groups, with the silk fibroincoated leaves again scoring the largest percentages with a 1 or 2 scoreat 95%. The control group and the water groups only had approximately89% of their leaves achieve this score. Finally, FIG. 10 shows rotscores, as a percentage of layers with a 1, across the various groups.In this experiment, the 100% of the silk fibroin coated layers had a rotscore of 1, representing that only 0-10% of the surface of the totesurface had rot. In contrast, the control and water groups only had rotscores of 1 on 67% of the tote layers.

These results exemplify the benefits of coating product with silkfibroin, as it decreases the negative effects of packing more productinto the same volume of container compared to other methods. It alsoshows that increased temperatures do not negatively impact theperformance of silk fibroin compared to other methods. This latter pointis very important because shipping and storage conditions are not alwaysuniform, and heat spikes can occur, either due to equipment issues ordelays in transportations. By reducing the harm to the product duringshipment disruptions, the silk fibroin coating provides flexibility toshippers and limits food waste. In another aspect, different scoringsystems could be created and analyzed to determine the impacts ofcoatings on a product.

The particulars of the study could be changed to create the desiredstress on the coated product. In one aspect, greater or fewer heatspikes could be used, for example from 1-5 heat spikes. In anotheraspect, the temperatures of the heat spikes could be modified, forexample, from 80-120 degrees Fahrenheit. In another aspect, the holdtimes of the heat spikes could also be modified, for example, from 15minutes to 1 hour per spike. In another aspect, the frequency of theheat spikes could also be modified by, for example, exposing the productto spikes every 12 hours or once every day. In another aspect, thetreatment of the product could also be altered to include non-silkfibroin coatings or multiple groups of different coatings forcomparison. In another aspect, different materials could be used to coatthe product. In another aspect, a different product entirely could beused, such as a different type of product.

Example 2 Shelf Life Study

A second model was developed to create conditions encountered duringtransport and storage of greens (e.g., spinach, kale, lettuce, etc.) totest the performance of silk fibroin coatings against uncoated greenspacked to normal tote weight, uncoated greens packed at a greater toteweight, and greens coated with silk fibroin packed at a greater toteweight. This experiment was designed to mimic a normal harvest to tabletimeline that consumers encounter, to measure the impact of silk fibroinon the greens. The increased packing weight was utilized to create astressed condition for the greens to study the impact of silk fibroinand confirm if greater packing densities were possible as part of theshipping/transport process.

In this exemplary experiment, the greens (baby kale) were collected by aharvester and immediately separated into three groups: the control group(untreated kale packed to 18 pounds); an untreated group (untreated kalepacked to 22 pounds): and a silk coated group (kale treated with silkfibroin solution and packed to 22 pounds). The silk fibroin was appliedvia spraying as the greens were harvested from the field by a spray bar,prior to packing the greens in totes. As mentioned, the control groupwas not treated and was packed to 18 pounds in totes. The untreatedgroup was not treated and packed to 22 pounds in totes. The silk fibroincoated group was treated with silk fibroin and packed to 22 pounds intotes. The totes were fully utilized at 22 pounds, such that noadditional kale could be added. FIG. 11 shows the different groups intheir totes on day 1 of the experiment.

The next step is to either transport the totes to the testing lab via atrailer (e.g., a refrigerated tractor trailer) or simulate the transporttime by storing the totes in the same type of cold storage for the sameamount of time it would take to physically transport the greens. In thisexperiment, the totes were physically transported by tractor trailer toCambridge Crops' lab over the course of approximately 5 days. Duringthis time the three groups were stored at temperatures of about 40degrees Fahrenheit and about 40% relative humidity. Once the totesarrived at Cambridge Crops, they were processed by being washed and thentransferred to clamshells to simulate the normal process fortransporting and then packing the greens for sale. Other containerscould be used besides clamshells, such as bags. These containers couldbe various sizes depending on the sale size intended. In this case, theclamshells were 5 ounces and the transported greens were evenly dividedup. For example, 13 clamshells were created for the control group, 13clamshells were created for the untreated group, and 14 clamshells werecreated for the silk coated group. FIG. 12 shows the different groupsdivided into their clamshells on day 9 of the experiment. FIG. 13 showsthe clamshells at day 28 of the experiment. The clamshells were thenstored in a cold room at 40 degrees Fahrenheit and 52% relative humidityfor the duration of the experiment. The experiment continued for 41 daysand the different clamshells were sampled throughout this period and atthe end of the experiment to provide data on the performance of eachgroup.

At the conclusion of the experiment the scores were complied todetermine if the groups differed throughout the experiment. Multipledifferent aspects of the product, in this case kale, can be observed andcompared, for example, metrics selected from one or more of visualinspection (e.g., yellowing, wilting, rot, color), structural integrity,microbial load (e.g., microbial growth), bounce back (e.g., the abilityof certain products (e.g., leafy greens) to recover from the negativeeffects of wilting once washed through rehydration, which shows assistsin showing whether the product was damaged) respiration rate, taste,temperature, insulative effect, UV filtering effect, and/or odor.

In this experiment wilting and yellowing were compared across thedifferent groups (e.g., control, untreated, silk coated). The groupswere compared by randomly selecting 15 leaves from the clamshells andassigning them a number based on the scales shown in FIGS. 1-2 . As canbe seen in FIG. 14 , the yellowing results over the first 28 days of theexperiment show that the untreated group is performing worse than theother two groups. This is more clearly shown in FIG. 15 , which showsthe percentage of leaves with a “good” yellowing score (meaning theyscored a 1). In this case, the control group and the silk treated grouphad the same percentage of leaves with a “good” score, whereas theuntreated group faired noticeably worse (about 10%). FIG. 16 shows thesame types of results, with the control group and the silk treatedgroups fairing better than the untreated group when both yellowing andwilting were examined. As this graph shows, a “good” score was achievedif yellowing was a 1 and wilting was a 1 or 2. Again, the control groupand the silk treated group outperformed the untreated group by day 28 byover 10%.

In this case, the silk coated group performed as well as the controlgroup despite being packed more tightly during transport. These resultsexemplify the benefits of coating produce, particularly kale, with silkfibroin, as it decreases the negative effects of packing more productinto the same volume of container compared to other methods. This willenable a shipper to add more kale to each shipment without worryingabout harming the kale by reducing its shelf life, which would not bethe case with untreated kale. This has the potential to save shippingcosts, reduce fuel consumption, remove trucks on the road, and forshippers to realize other efficiencies. In addition, the coating waswashed off the kale during the processing step. This showcases theknock-on effect of the silk coating, because it does not have to remainon the product to impact shelf life.

The study described above was exemplary and the particulars of the studycould be changed to create the desired stress on the coated product. Forexample, different materials could be used to coat the product. Inanother aspect, a different product entirely could be used, such as adifferent type of product. Different conditions could be implemented,such as by changing temperatures, humidity, duration, packing method,etc. Finally, different measurements of the product tested could betaken, such as respiration rate, structural integrity, microbial load,and the like.

Example 3 Respiration Testing

To study the respiration of different products, two different methodshave been developed. The first tests small sample sizes, generallybetween 20-50 grams that are placed in a jar (e.g., a mason jar). FIG.17 shows an example of this method, with the container (C), product (P),and testing device (F) shown. In this process the product (P) is placedin the contained (C) and an initial carbon dioxide (CO₂) measurement istaken. The container is then left to rest for a predetermined amount oftime (e.g., 15 minutes to an hour) and a second CO₂ measurement istaken. In this case, a Felix Instruments F-950 (F) was used to measurethe CO₂. A final rate of respiration is provided and is expressed as mLCO₂ kg⁻¹ hr⁻¹. In the second method, a larger container is tested (e.g.,a tote) for respiration rate. FIG. 18 shows the container (C) andtesting device (F). The same process is used as for FIG. 17 , just on alarger scale. The end result is another respiration rate measurementrepresented in mL CO₂ kg⁻¹ hr⁻¹.

1.-21. (canceled)
 22. A method of increasing the performance of atransport system having a performance variable, wherein the transportsystem transports, stores a product, or both, and wherein the productcomprises at least one of edible proteins, fruits, vegetables, nuts,seeds, flowers, plants, pet food, animal feed, or combinations thereof,the method comprising: coating at least part of the surface of anuncoated product with a substance to create a coated product, whereinthe substance is selected from the group of: silk fibroin, chitin,acetylated monoglycerides, shellac, starch, high fructose corn syrup,mayauba wax, maydelilla wax, beeswax, vegetable oil, paraffin oil,ethylenediaminetetraacetic acid, cellulose, pectin, alginate, chitosan,gum Arabic, soy protein, zein, casein, whey, and combinations thereof;thereby improving at least one performance variable of the coatedproduct compared to the uncoated product when the product is at leastone of transported and stored in the transport system.
 23. The method ofclaim 22, further including at least one of a step of transporting and astep of storing the coated product in the transport system for a periodof time.
 24. The method of claim 22, further comprising a step ofincreasing the freight capacity of the transport system, wherein thetransport system has a first freight capacity of the product prior tocoating and a second freight capacity of the coated product, and whereinthe second freight capacity is larger than the first freight capacity.25. The method of claim 24, wherein the freight capacity of the systemis the weight of the product, and wherein the second freight capacity isincreased by at least 5% compared to the first freight capacity.
 26. Themethod of claim 25, wherein the second freight capacity is increased byat least 15% compared to the first freight capacity.
 27. The method ofclaim 24, wherein the freight capacity of the system is the volumeutilization of the transport system, and wherein the second freightcapacity is increased by at least 5% compared to the first freightcapacity.
 28. The method of claim 22, wherein the product is coatedpre-harvest.
 29. The method of claim 22, wherein the product is coatedpost-harvest.
 30. The method of claim 22, wherein the product is coatedduring harvest.
 31. The method of claim 30, wherein the product iscoated by a harvester.
 32. The method of claim 29, wherein the productcomprises produce and the product is coated when the produce is washed.33. The method of claim 22, wherein the product is coated during atleast one of transportation and a distribution process.
 34. The methodof claim 22, wherein at least one of the performance variable(s) of theproduct is selected from one of the following: color, structuralintegrity, respiration rate, microbial load, yellowing, wilting, bounceback, taste, odor, temperature, insulative effect, UV filtering effect,and rot.
 35. The method of claim 34, wherein yellowing of the product isreduced by 5%.
 36. The method of claim 34 wherein wilting of the productis reduced by 5%.
 37. The method of claim 34, wherein rot of the productis decreased by 5%.
 38. The method of claim 22, wherein the productcomprises one of spinach, kale, arugula, lettuce, spring mix, cilantro,broccoli, green onion, and parsley.